The Remodeling Of Gap Junctional Protein In Vivo Experiments Of Cerebral Vasospasm

Objective: Subarachnoid hemorrhage(SAH) is one of the common but severe cerebrovascular accidents. Cerebral vasospasm(CVS) is the major dangerous factor induced by SAH. The mechanism of CVS is still unknown, and there haven't effective method to treat it till now. Gap junctions(GJ), which serve as channels for direct intercellular exchange of ions, and small signaling molecules, play an important role in communication between adjacenct cells. We have indicated that GJ are involved in CVS during early phase. The main objective of this study is to explore the change of expression of connexin43(Cx43) mRNA, Cx43 protein and its distribution after the model of subarachnoid two hemorrhage in rabbits, which will provide the basis to study the mechanism of CVS and its genetic treatment.METHODS: The model of subarachnoid two hemorrhage in New Zealand rabbits was established. The expressiong of Cx43 mRNA of basilar arteries tissue in rabbits was detected by RT-PCR. The rabbits were grouped as control and models(included 24h, 48h, 72h,96h and 7d groups after two hemorrhage).The expressiong of Cx43 of basilar arteries tissue was examined by using western blotting analysis. Using immunohistochemical technique, the location of Cx43 in the wall of basilar artery was evaluated.RESULT: The model of subarachnoid two hemorrhage in rabbits was successfully established. The results of semiquantitative RT-PCR showed that the level of Cx43 mRNA in models increased gradually and significantly compared with that of control(P<0.01), the expression of 72h group was the highest, and then looks lower gradually. The expression of Cx43 by western blotting analysis was similar to the level of Cx43 mRNA by RT-PCR. Immunohistochemistry indicated that the Cx43 was found in the endothelial and outer membrane layer in control, but not distributed in the smooth muscle layer of the basilar artery. Our results showed interestedly that the Cx43 was located in the smooth muscle layer in the models. CONCLUSION: The above results demonstrated for the first time that the connexin was remodeled after SAH, which might be connected with the development of CVS. Our study may provide insight into the explanation for CVS and the new genetic drug development strategy for preventing and treating it.